维生素E脂质体纳米颗粒转运siRNA抑制丙型肝炎病毒核心蛋白表达的实验研究

发布时间：2018-03-03 21:55

本文选题：丙型肝炎　切入点：RNA干扰　出处：《第三军医大学学报》2017年20期 　论文类型：期刊论文

【摘要】：目的研究携带针对丙型肝炎病毒(hepatitis C virus,HCV)RNA的小干扰RNA(siRNA)的维生素E(Vitamin E)脂质体纳米颗粒在体外靶向抑制HCV复制的作用。方法采取"薄膜水化法"制备Vitamin E脂质体纳米颗粒,应用激光粒度分析仪测量纳米颗粒VE-DC/siRNA的颗粒大小及zeta电位,应用透射电子显微镜观察VE-DC形态及分散性。以Huh7.5.1细胞株作为载体。CCK-8法检测纳米颗粒的细胞毒性,琼脂糖凝胶电泳检测纳米颗粒携带siRNA的血清稳定性,蛋白印迹技术和免疫荧光技术检测该siRNA纳米颗粒的转染效率及对HCV的抑制作用。结果 Vitamin E脂质体纳米颗粒形态呈球形,粒度分布为(125.5±9.0)nm,zeta电位为(39.1±4.8)m V,分散均一。相较同浓度商品化脂质体,VE-DC/siRNA对细胞的毒性相对较小。血清稳定性实验显示,VE-DC/siRNA在24 h后siRNA无降解,较裸siRNA稳定性显著提高。携带siRNA的Vitamin E脂质体纳米颗粒可以较高效率进入Huh7.5.1细胞内,并抑制HCV核心蛋白的表达。结论 Vitamin E脂质体纳米颗粒可以转运siRNA至Huh7.5.1细胞,并抑制HCV核心蛋白的表达。
[Abstract]:Objective to study the inhibition of HCV replication by liposome nanoparticles carrying small interfering RNAs against hepatitis C virus (HCV) virus. Methods Vitamin E liposome nanoparticles were prepared by "membrane hydration method". The particle size and zeta potential of VE-DC/siRNA nanoparticles were measured by laser particle size analyzer, and the morphology and dispersion of VE-DC were observed by transmission electron microscope. The cytotoxicity of nanoparticles was detected by using Huh7.5.1 cell line as carrier. CCK-8 method was used to detect the cytotoxicity of nanoparticles. Agarose gel electrophoresis was used to detect the serum stability of the nanoparticles carrying siRNA, Western blot and immunofluorescence techniques were used to detect the transfection efficiency of the nanoparticles and their inhibitory effect on HCV. Results the Vitamin E liposome nanoparticles were spherical in shape. The particle size distribution was 125.5 卤9.0nmnmdeta potential (39.1 卤4.8mV). The cytotoxicity of VE-DCR siRNA was relatively less than that of commercial liposome at the same concentration. Serum stability test showed that VE-DCR / siRNA did not degrade after 24 h. Vitamin E liposome nanoparticles carrying siRNA could enter Huh7.5.1 cells more efficiently and inhibit the expression of HCV core protein. Conclusion Vitamin E liposome nanoparticles can transport siRNA to Huh7.5.1 cells. And inhibit the expression of HCV core protein.
【作者单位】：重庆医科大学附属第二医院科研处;重庆医科大学附属第二医院检验科;重庆医科大学附属第二医院感染科;
【基金】：国家自然科学基金面上项目(81171628)~~
【分类号】：R450;R512.63

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